CN108835385B - Compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns - Google Patents

Compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns Download PDF

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CN108835385B
CN108835385B CN201810867168.0A CN201810867168A CN108835385B CN 108835385 B CN108835385 B CN 108835385B CN 201810867168 A CN201810867168 A CN 201810867168A CN 108835385 B CN108835385 B CN 108835385B
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chinese herbal
prawn
herbal medicine
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detoxification
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CN108835385A (en
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潘鲁青
苏晨
路玉松
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Ocean University of China
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/20—Animal feeding-stuffs from material of animal origin
    • A23K10/22—Animal feeding-stuffs from material of animal origin from fish
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/111—Aromatic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/116—Heterocyclic compounds
    • A23K20/121—Heterocyclic compounds containing oxygen or sulfur as hetero atom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish
    • Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

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Abstract

A compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns comprises four Chinese herbal medicines of radix bupleuri, fructus forsythiae, eucommia ulmoides and liquorice, wherein the mass ratio of the active ingredients of the Chinese herbal medicines is as follows: 20-40% of saikosaponin, 20-40% of forsythin, 10-30% of liquorice and 10-30% of chlorogenic acid. After the preparation is produced and applied, the activity of the prawn hepatopancreas detoxification metabolizing enzyme and the antioxidation level can be obviously improved, the detoxification capacity of the prawn is promoted, and the normal function of the hepatopancreas is protected. The invention extracts four Chinese herbal medicine active ingredients as the compound Chinese herbal medicine detoxification liver-protecting regulator, has the characteristics of simple preparation, low cost, definite active ingredients, high curative effect and the like, and provides technical support for guaranteeing the physiological health of the prawns.

Description

Compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns
Technical Field
The invention relates to a compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns, belonging to the technical field of aquatic compound feed additives.
Background
With the rapid development of the shrimp culture industry in China, the obvious problems of difficult disease prevention and control, misuse of fishing drugs, excessive and unbalanced nutrition in feed and the like occur due to blind pursuit of a culture mode with rapid growth and high yield, the detoxification metabolism of the prawns is blocked, the physiological health level is reduced, and the green and healthy development of the shrimp culture industry is severely restricted.
The hepatopancreas is the most important organ of the prawn body and plays a very key physiological role. The existing prawn compound feed has the problem that protein and fat are excessively added to cause the accumulation of the hepatopancreas fat of the prawn. Meanwhile, the increase of plant proteins in the feed instead of animal proteins can cause that the feed is very easy to breed mould to generate cancerogenic and mutagenic substances such as aflatoxin B1 and the like, and cause that the metabolism of liver pancreas is disturbed and endotoxin is increased. In order to prevent the loose-leaf disease outbreak of the culture pond, a large amount of chemical substances such as antibiotics and the like can bring heavy burden to the hepatopancreas of the prawn, so that the physiological health and industrial sustainable development of the prawn are seriously damaged, and the maintenance of the hepatopancreas of the prawn becomes an urgent research subject to be solved.
At present, researches on prawn feed additives are focused on aspects of enhancing organism immunity, disease resistance and the like of prawns, such as enhancing nonspecific immunity of prawns by adding immune enhancing agents such as immune polysaccharide, chinese herbal medicines and the like, and researches on protecting liver and pancreas of prawns are shallow. Meanwhile, the traditional Chinese herbal medicine additive is mainly prepared by crushing medicines and then directly adding the crushed medicines into feed, has the problems of undefined active ingredients, more impurities, poor palatability and the like, and is easy to cause hepatopancreatic injury and cultivation environment pollution.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention mainly aims to provide the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns so as to enhance the detoxification metabolic capacity of the liver and pancreas of the prawns. The preparation adopts four Chinese herbal medicines, extracts the effective components of the Chinese herbal medicines, and adds the effective components into feed, so that the antioxidant activity and the detoxifying metabolic enzyme activity of the hepatopancreas of the prawn are enhanced, and the added components are green and safe and can not pollute the culture water body.
In order to achieve the purpose, the invention adopts the following specific technical scheme:
a compound Chinese herbal medicine feed additive for detoxication and liver protection of prawns is characterized by comprising 20-40% of saikosaponin, 20-40% of forsythin, 10-30% of liquorice and 10-30% of chlorogenic acid by mass fraction.
The compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns is characterized by comprising saikosaponin: forsythin: chlorogenic acid: the mass ratio of glycyrrhizic acid is 5:5:5:3.
the compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns is characterized by comprising saikosaponin: forsythin: chlorogenic acid: the mass ratio of glycyrrhizic acid is 5:5:2:3.
the saikosaponin is extracted from bupleurum, the forsythin is extracted from forsythia, the chlorogenic acid is extracted from eucommia ulmoides, and the glycyrrhizic acid is extracted from liquorice. Extracting saikosaponin and forsythin with water, extracting chlorogenic acid with ethyl acetate, and extracting glycyrrhizic acid with diluted ammonia water.
The saikosaponin, the forsythin, the chlorogenic acid and the glycyrrhizic acid are prepared by the following steps:
the saikosaponin and the forsythin are prepared by respectively crushing the bupleurum and the forsythia, respectively mixing with distilled water according to the mass-volume ratio of 1:10, decocting for 2 times, each time for 2 hours, combining 2 times of filtrate, concentrating under reduced pressure, and freeze-drying to obtain the saikosaponin and the forsythin;
the chlorogenic acid is prepared by uniformly crushing flaky eucommia ulmoides, uniformly mixing the crushed flaky eucommia ulmoides with ethyl acetate containing 0.05mol/L hydrochloric acid according to a mass-volume ratio of 1:10, stirring, refluxing and leaching for 2 hours on a water bath at 65 ℃, repeatedly extracting for 2 times, mixing the filtrates, and performing rotary evaporation;
extracting glycyrrhizic acid with dilute ammonia water, pulverizing Glycyrrhrizae radix, extracting with 0.5% dilute ammonia water twice for 2 hr each time, mixing the filtrates, adding 98% concentrated sulfuric acid, stirring thoroughly to adjust pH to 2, standing, filtering, washing with water for 2-3 times, and lyophilizing to obtain glycyrrhizic acid.
The compound prawn Chinese herbal medicine detoxification liver-protecting feed additive can be applied to preparation of prawn culture feeds, and can enhance the detoxification metabolism capacity of the liver pancreas of prawns.
A prawn feed containing prawn compound Chinese herbal medicine detoxification liver protection feed additive is characterized in that the prawn compound Chinese herbal medicine detoxification liver protection feed additive is dissolved in water and then uniformly sprayed on the surface of basic feed, fish oil is used for wrapping the feed, and the prawn feed containing the prawn compound Chinese herbal medicine detoxification liver protection feed additive is obtained after airing.
The spraying amount of the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns in each kilogram of basic feed is as follows: 0.1g of saikosaponin, 0.1g of forsythin, 0.1g of chlorogenic acid and 0.06g of glycyrrhizic acid; or 0.25g of saikosaponin, 0.25g of forsythin, 0.1g of chlorogenic acid and 0.15g of glycyrrhizic acid.
The basic feed comprises 34% of fish meal, 20% of soybean meal, 16.4% of peanut meal, 22% of wheat flour, 5% of soybean lecithin, 0.6% of compound vitamin and 2% of compound mineral substances in percentage by mass.
The invention has the advantages and beneficial effects that:
(1) The total antioxidant capacity of the litopenaeus vannamei is improved. The total antioxidant capacity index represents the total antioxidant activity of all antioxidant substances of the organism including enzymes and nonenzymes. Within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive for the prawns and 10 days of the aflatoxin B1 toxicity test, the T-AOC of the prawns in the test group is obviously higher than that of the prawns in the control group (P < 0.05).
(2) The superoxide dismutase activity of the litopenaeus vannamei is improved. Superoxide dismutase is the first antioxidant enzyme to act in the active oxygen scavenging system. Within 28 days of feeding the compound Chinese herbal medicine detoxification liver protection feed additive for prawns and 10 days of the aflatoxin B1 toxicity test, the SOD activity of the prawns in the test group is obviously higher than that of the prawns in the control group (P < 0.05).
(3) The content of cytochrome P450 of the litopenaeus vannamei is improved. Cytochrome P450 (CYP 450) catalyzes the monooxygenating reaction of endogenous and exogenous compounds in phase I metabolism. Within 28 days of feeding the compound Chinese herbal medicine detoxification liver protection feed additive for prawns and 10 days of the aflatoxin B1 toxicity test, the CYP450 content of the test group prawns is obviously higher than that of the control group P < 0.05).
(4) The activity of glutathione-transferase (GST) of litopenaeus vannamei is improved. Glutathione Sulfhydryl Transferase (GST) is one of the most important phase II metabolic enzymes for in vivo biotransformation, and is a main detoxification system for cell damage resistance and canceration resistance. The GST activity of the test group is obviously higher than that of the control group P <0.05 within 28 days of feeding the compound Chinese herbal medicine detoxification liver protection feed additive for prawns and 10 days of the toxicity test of aflatoxin B1.
(5) In the toxicity test of aflatoxin B1, the activity of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase in the blood plasma of litopenaeus vannamei is reduced. Glutamic pyruvic transaminase and glutamic oxaloacetic transaminase activities are the most sensitive indicators of liver cell damage, and can evaluate the health degree of liver pancreas. Within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns, the activity of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase of a control group and a test group is not obviously different (P is more than 0.05), and within 10 days of an aflatoxin B1 toxicity test, the activity of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase of the test group is obviously lower than that of the control group (P is less than 0.05).
Experiments prove that the prawn compound Chinese herbal medicine detoxification liver-protecting feed additive provided by the invention can improve the detoxification metabolism capability and the antioxidation capability of prawn pancreas, and the enhancer is prepared from four Chinese herbal medicines of bupleurum, weeping forsythiae capsule, eucommia bark and liquorice effectively and scientifically.
Drawings
FIG. 1 is a graph showing the results of the total antioxidant capacity of the hepatopancreas of prawns in example 2.
FIG. 2 is a graph showing the results of the activity of the hepatopancreatic superoxide dismutase of example 2.
FIG. 3 is a graph showing the results of the content of the hepatopancreatic cytochrome P450 of prawn in example 2.
FIG. 4 is a graph showing the results of the activity of the thiol transferase of hepatopancreatic glutathione of example 2.
FIG. 5 is a graph showing the results of the activity of glutamic pyruvic transaminase in the plasma of prawn in example 2.
FIG. 6 is a graph showing the results of the activity of glutamic-oxaloacetic transaminase in the plasma of prawn in example 2.
Description of the embodiments
Example 1:
compound Chinese herbal medicine feed additive for detoxicating and protecting liver of prawn and preparation of prawn feed
Firstly, respectively crushing bupleurum and weeping forsythiae capsule, uniformly mixing the crushed bupleurum and weeping forsythia capsule with distilled water according to the mass volume ratio of 1:10, decocting for 2 times, each time for 2 hours, combining 2 times of filtrate, concentrating under reduced pressure, and drying to obtain the saikosaponin and the weeping forsythin capsule; uniformly pulverizing sheet eucommia ulmoides, uniformly mixing with ethyl acetate (containing 0.05mol/L hydrochloric acid) according to a mass-volume ratio of 1:10, stirring, refluxing and leaching for 2 hours on a 65 ℃ water bath, repeatedly extracting for 2 times, mixing filtrates, and rotationally evaporating to obtain chlorogenic acid; extracting glycyrrhizic acid with dilute ammonia water, pulverizing Glycyrrhrizae radix, extracting with 0.5% dilute ammonia water twice for 2 hr each time, mixing the filtrates, adding concentrated sulfuric acid, stirring thoroughly to adjust pH to 2, washing with water, filtering, and drying to obtain glycyrrhizic acid.
The Chinese herbal medicine extract is dissolved in water and then uniformly sprayed on the surface of the basic feed, the feed is wrapped by fish oil, and the basic feed added with the compound Chinese herbal medicine detoxification liver-protecting feed additive of the prawns is obtained after airing.
Two reinforcing agents with different concentrations and corresponding breeding feeds are respectively prepared according to the preparation method. The compound Chinese herbal medicine feed additive for detoxifying and protecting liver of the prawns comprises, by mass, 0.1g of saikosaponin, 0.1g of forsythin, 0.1g of chlorogenic acid and 0.06g of glycyrrhizic acid added into each kilogram of feed; the other compound Chinese herbal medicine feed additive for detoxifying and protecting liver of prawns comprises 0.25g of saikosaponin, 0.25g of forsythin, 0.1g of chlorogenic acid and 0.15g of glycyrrhizic acid added into each kilogram of feed.
The content of the effective components of the above-mentioned Chinese herbal medicine can be measured according to the methods provided in the prior art. The method for measuring the content of the saikosaponin is as follows: measuring the contents of total saponins and saikosaponin a in bupleurum in different producing areas, and measuring the contents of Chinese medicines and clinical medicines in volume 8 and 3 of 2008; the method for measuring the content of the forsythin is as follows: content analysis of forsythoside and forsythoside in different parts of fructus forsythiae, journal of pharmaceutical analysis, volume 15, 3 of 2008; the method for measuring chlorogenic acid content refers to: journal of Pharmaceutical Analysis, 2011, volume 1, 04; the method for measuring the glycyrrhizic acid content refers to the following steps: the quantitative method of glycyrrhizic acid in Glycyrrhrizae radix and its preparation is studied and outlined, and the medicine of Shizhen national medicine, volume 11, 4 in 2000.
Examples
Influence of compound Chinese herbal medicine detoxification liver-protecting feed additive of prawns on oxidation resistance and detoxification metabolism capacity of litopenaeus vannamei
In the test, the inventor feeds the compound prawn Chinese herbal medicine detoxification liver protection feed additive prepared in the example 1 to litopenaeus vannamei, and researches the effect of improving the detoxification metabolism capacity of the litopenaeus vannamei hepatopancreas by analyzing the detoxification metabolism and antioxidation indexes of the litopenaeus vannamei.
1. Materials and methods
1.1 Material
Litopenaeus vannamei used for testLitopenaeus vannamei) The feed is purchased from a shrimp culture factory in Laoshan mountain area in Qingdao city, and has the weight of 3+/-0.32 g, and the ingredients of the basic feed for the test are as follows: 34% of fish meal, 20% of soybean meal, 16.4% of peanut meal, 22% of wheat flour, 5% of soybean lecithin, 0.6% of multivitamin and 2% of compound mineral substances. The experimental calculation shows that the feed comprises the following nutritional components: the mass fraction of the crude protein is 42.4%, the mass fraction of the crude fat is 7.2% and the mass fraction of the ash is 8.4%.
1.2 Method of
1.2.1 Test design
The test prawns were divided into 3 groups of 3 replicates each, each replicate group stocking 80 prawns. The test components are as follows: control group, test group 1 and test group 2, wherein the control group is fed with basic feed (added with fish oil with the same amount as that of the prawns in the test group), and the test groups 1 and 2 are respectively fed with the compound Chinese herbal medicine detoxification liver-protecting feed additive feed containing the prawns and prepared in the example 1. The cultivation experiment lasts for 28 days. After the cultivation experiment is finished, a toxicity test is carried out, and aflatoxin (2500 mug/kg) is added to the control group and the test group on the basis of the original feed, wherein the toxicity test lasts for 10 days. During the test, the feed is fed for 2 times a day, the daily feed amount is 5% of the weight, and during the test, the water temperature, the salinity and the PH are respectively 22-23 ℃, 34 per mill and 7.8-8.0.
1.2.2 Measurement index and method
Sampling is carried out on days 0, 7, 14, 21, 28, 31, 34 and 38 after feeding the compound Chinese herbal medicine detoxification liver protection feed additive of the prawns. Each group randomly taking 6 litopenaeus vannamei, directly inserting the litopenaeus vannamei into the heart chamber around 3mm at the rear edge of the chest nail of the prawn by using a sterilized 5-gauge needle and a 1mL syringe to collect blood, and pre-sucking 0.3mL of improved precooled litopenaeus vannamei in the syringe before blood collectionAnti-coagulant for litopenaeus (0.34M NaCl, 0.01M KCl, 0.01M EDTA-Na) 2 0.01M HEPES, pH 7.45, osmolality 780 mOsm.kg -1 ) Finally, the ratio of anticoagulant to haemolymph is 1:1. After centrifugation of 800g of the haemolymph sample at 4 ℃ for 10min, the blue supernatant was taken and was the plasma sample. Dissecting and taking hepatopancreatic samples, placing the hepatopancreatic samples into a mortar, grinding the hepatopancreatic samples by liquid nitrogen, and preserving all the samples in a refrigerator at the temperature of-80 ℃.
T-AOC was determined using Nanjing's built biosystems production kit; measurement of superoxide dismutase activity is carried out according to the methods of the prior art, and reference can be made to: involvement of The superoxide anion radical in The autoxidation of pyrogallol and a convenient assay for superoxide The FEBS Journal; the measurement of cytochrome P450 activity was carried out according to the methods of the prior art, and reference may be made to: the carbon monoxide-binding pigment of liver microsomes I Evidence for its hemoprotein natural Journal of Biological Chemistry; the measurement of glutathione thiol transferase activity was performed according to the methods of the prior art, and can be referred to as follows: glutethione S-transferases the first enzymatic step in mercapturic acid information Journal of biological Chemistry; glutamic pyruvic transaminase and glutamic oxaloacetic transaminase are determined using a kit for manufacturing Nanjing built biosystems.
2. Results
2.1 Total antioxidant Activity
As can be seen from fig. 1: within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns, the total antioxidant capacity of the prawns in the test group is obviously higher than that in the control group (P < 0.05), the total antioxidant capacity is highest on the 7 th day of feeding, but the difference between the test group 1 and the test group 2 is not obvious (P > 0.05), the total antioxidant capacity of the control group is reduced in the aflatoxin B1 toxicity test, and the total antioxidant capacity of the test group is obviously higher than that of the control group (P < 0.05).
2.2 Superoxide dismutase activity
As can be seen from fig. 2: within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns, the superoxide dismutase of the prawns in the test group is obviously higher than that in the control group (P < 0.05), the SOD activity on the 7 th day and the SOD activity on the 14 th day are higher, and the difference between the test group 1 and the test group 2 on the 28 th day is not obvious (P > 0.05). In the aflatoxin B1 toxicity test, the total antioxidant capacity of a control group is greatly reduced, the SOD activity of a test group is obviously higher than that of the control group (P < 0.05), the SOD activity of a test group 2 is higher than that of a test group 1, and the difference between the test groups is not obvious (P > 0.05).
2.3 CYP450 content
As can be seen from fig. 3: within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns, the CYP450 content of the prawns in the test group is obviously higher than that of the control group (P < 0.05), the CYP450 content of the test group 2 on the 7 th day and the 21 st day is obviously higher than that of the test group 1 (P < 0.05), the CYP450 content of the control group and the test group is not obviously reduced in the aflatoxin B1 toxicity test, the CYP450 content of the test group tends to be stable, and the difference is not obvious (P > 0.05).
2.4 Glutathione thiol transferase Activity
As can be seen from fig. 4: within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive to the prawns, the activity of the hepatopancreatic glutathione sulfhydryl transferase of the prawns is obviously higher than that of a control group (P < 0.05), the GST activity of the test group reaches the highest within 28 days, and the difference between the test group 2 and the test group 1 is obvious (P < 0.05). In the aflatoxin B1 toxicity assay, the control GST activity was greatly reduced, and the test GST activity was significantly higher than the control (P < 0.05).
2.5 Glutamic pyruvic transaminase activity
As can be seen from fig. 5: within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive for prawns, the activity of the plasma glutamic pyruvic transaminase of the prawns in the test group and the control group is not obviously different (P is more than 0.05), which indicates that the Chinese herbal medicine detoxification liver-protecting regulator has no damage effect on the litopenaeus vannamei. In the aflatoxin B1 toxicity test, the glutamic-pyruvic transaminase activity of a control group is greatly increased, the glutamic-pyruvic transaminase activity of a test group is obviously lower than that of the control group (P < 0.05), and the difference among the test groups is not obvious (P > 0.05) along with the duration of the toxicity test.
2.6 Glutamic-oxaloacetic transaminase activity
As can be seen from fig. 6: within 28 days of feeding the compound Chinese herbal medicine detoxification liver-protecting feed additive to the prawns, the activity of the plasma glutamic-oxaloacetic transaminase of the prawns in the test group and the control group is not obviously different (P is more than 0.05). In the aflatoxin B1 toxicity test, the activity of the glutamic-oxaloacetic transaminase in the plasma of a control group is greatly increased, the activity of the glutamic-oxaloacetic transaminase in the test group is obviously lower than that of the control group (P < 0.05), and the difference among the test groups is obvious (P < 0.05) along with the duration of the toxicity test.

Claims (7)

1. A compound Chinese herbal medicine feed additive for detoxication and liver protection of prawns is characterized by comprising 20-40% of saikosaponin, 20-40% of forsythin, 10-30% of glycyrrhizic acid and 10-30% of chlorogenic acid by mass fraction,
the saikosaponin, the forsythin, the chlorogenic acid and the glycyrrhizic acid are prepared by the following steps:
the saikosaponin and the forsythin are prepared by respectively crushing the bupleurum and the forsythia, respectively mixing with distilled water according to the mass-volume ratio of 1:10, decocting for 2 times, each time for 2 hours, combining 2 times of filtrate, concentrating under reduced pressure, and freeze-drying to obtain the saikosaponin and the forsythin;
the chlorogenic acid is prepared by uniformly crushing flaky eucommia ulmoides, uniformly mixing the crushed flaky eucommia ulmoides with ethyl acetate containing 0.05mol/L hydrochloric acid according to a mass-volume ratio of 1:10, stirring, refluxing and leaching for 2 hours on a water bath at 65 ℃, repeatedly extracting for 2 times, mixing the filtrates, and performing rotary evaporation;
extracting glycyrrhizic acid with dilute ammonia water, pulverizing Glycyrrhrizae radix, extracting with 0.5% dilute ammonia water twice for 2 hr each time, mixing the filtrates, adding 98% concentrated sulfuric acid, stirring thoroughly to adjust pH to 2, standing, filtering, washing with water for 2-3 times, and lyophilizing to obtain glycyrrhizic acid.
2. The compound Chinese herbal medicine feed additive for prawn detoxification and liver protection according to claim 1, wherein the saikosaponin: forsythin: chlorogenic acid: the mass ratio of glycyrrhizic acid is 5:5:5:3.
3. the compound Chinese herbal medicine feed additive for prawn detoxification and liver protection according to claim 1, wherein the saikosaponin: forsythin: chlorogenic acid: the mass ratio of glycyrrhizic acid is 5:5:2:3.
4. use of the compound prawn Chinese herbal medicine detoxification liver protection feed additive according to any one of claims 1-3 in preparing prawn cultivation feed.
5. A prawn feed containing a prawn compound Chinese herbal medicine detoxification liver protection feed additive is characterized in that the prawn compound Chinese herbal medicine detoxification liver protection feed additive of claim 1 is dissolved in water, then uniformly sprayed on the surface of a basic feed, and then the feed is wrapped by fish oil, and finally dried to obtain the prawn feed containing the prawn compound Chinese herbal medicine detoxification liver protection feed additive.
6. The prawn feed of claim 5, wherein the compound Chinese herbal medicine detoxification and liver protection feed additive of prawn in each kilogram of basic feed has the following spraying amount: 0.1g of saikosaponin, 0.1g of forsythin, 0.1g of chlorogenic acid and 0.06g of glycyrrhizic acid; or 0.25g of saikosaponin, 0.25g of forsythin, 0.1g of chlorogenic acid and 0.15g of glycyrrhizic acid.
7. The prawn feed of claim 5 or 6, wherein the basal feed comprises, by mass, 34% of fish meal, 20% of soybean meal, 16.4% of peanut meal, 22% of wheat flour, 5% of soybean lecithin, 0.6% of compound vitamin and 2% of compound mineral.
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